Phospho-selective mechanisms of arrestin conformations and functions revealed by unnatural amino acid incorporation and (19)F-NMR

• Published in: Nature communications 2015-09, Vol.6, p.8202
• Main Authors: Yang, Fan, Yu, Xiao, Liu, Chuan, Qu, Chang-Xiu, Gong, Zheng, Liu, Hong-Da, Li, Fa-Hui, Wang, Hong-Mei, He, Dong-Fang, Yi, Fan, Song, Chen, Tian, Chang-Lin, Xiao, Kun-Hong, Wang, Jiang-Yun, Sun, Jin-Peng
• Format: Article
• Language: English
• Subjects: Animals; Arrestins - genetics; Arrestins - metabolism; beta-Arrestin 1; beta-Arrestins; Binding Sites; Blotting, Western; Cattle; Clathrin - metabolism; Escherichia coli; Fluorine; Fluorine-19 Magnetic Resonance Imaging; G-Protein-Coupled Receptor Kinase 2 - metabolism; G-Protein-Coupled Receptor Kinases - metabolism; HEK293 Cells; Humans; Microscopy, Confocal; Mutation; Nuclear Magnetic Resonance, Biomolecular; Phosphate-Binding Proteins - metabolism; Phosphoproteins - metabolism; Protein Conformation; Receptors, G-Protein-Coupled - metabolism; Signal Transduction; Tandem Mass Spectrometry; Tyrosine - analogs & derivatives; Tyrosine - metabolism
• ISSN: 2041-1723
• DOI: 10.1038/ncomms9202

Specific arrestin conformations are coupled to distinct downstream effectors, which underlie the functions of many G-protein-coupled receptors (GPCRs). Here, using unnatural amino acid incorporation and fluorine-19 nuclear magnetic resonance ((19)F-NMR) spectroscopy, we demonstrate that distinct receptor phospho-barcodes are translated to specific β-arrestin-1 conformations and direct selective signalling. With its phosphate-binding concave surface, β-arrestin-1 'reads' the message in the receptor phospho-C-tails and distinct phospho-interaction patterns are revealed by (19)F-NMR. Whereas all functional phosphopeptides interact with a common phosphate binding site and induce the movements of finger and middle loops, different phospho-interaction patterns induce distinct structural states of β-arrestin-1 that are coupled to distinct arrestin functions. Only clathrin recognizes and stabilizes GRK2-specific β-arrestin-1 conformations. The identified receptor-phospho-selective mechanism for arrestin conformation and the spacing of the multiple phosphate-binding sites in the arrestin enable arrestin to recognize plethora phosphorylation states of numerous GPCRs, contributing to the functional diversity of receptors.